Reverse transcription polymerase chain reaction

In molecular biology, reverse transcription polymerase chain reaction (RT-PCR) is a laboratory technique for amplifying a defined piece of a ribonucleic acid (RNA) molecule. The RNA strand is first reverse transcribed into its DNA complement or complementary DNA, followed by amplification of the resulting DNA using polymerase chain reaction. This can either be a 1 or 2 step process.

Polymerase chain reaction itself is the process used to amplify specific parts of a DNA molecule, via the temperature-mediated enzyme DNA polymerase.

Reverse transcription PCR is not to be confused with real-time polymerase chain reaction which is also marketed as RT-PCR.

Usage of reverse transcription polymerase chain reaction

The exponential amplification via reverse transcription polymerase chain reaction provides for a highly sensitive technique, where a very low copy number of RNA molecules can be detected. Reverse transcription polymerase chain reaction is widely used in the diagnosis of genetic diseases and, semiquantitatively, in the determination of the abundance of specific different RNA molecules within a cell or tissue as a measure of gene expression. Northern blot is used to study the RNA's gene expression further.

ee also

*Reverse transcriptase PCR

External links

* [ RT-PCR protocols from Penn state University]
* [ Database of validated PCR primer sets]
* [ Java Applet for testing rt-PCR primers]
* Visit the next international real-time PCR Event
* [ Animation to illustrate RT-PCR procedure, from Cold Spring Harbor Laboratory]

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